Havlas V, Kos P, Jendelová P, Lesný P, Trč T, Syková E
Ortopedická klinika UK, LF a FN Motol, dětská a dospělá ortopedie a traumatologie, Praha.
Acta Chir Orthop Traumatol Cech. 2011;78(2):138-44.
Many congenital and acquired disorders as well as sequelae of injury are associated with articular cartilage degeneration, which adversely affects the patient's quality of life. The currently used cell therapy with cultured chondrocytes has its disadvantages due to a process of de-differentiation of chondrocytes during cultivation. We believe that the mesenchymal stem cell therapy offers a new treatment options.
The adult mesenchymal stem cells (MSCs) for chondrocyte differentiation are usually obtained from bone marrow mesenchymal stem cells (BMSCs). In this study these cells were compared with mesenchymal stem cells derived from adipose tissue (AMSCs). The aim of the study was to verify the ability of human BMSCs and AMSCs to differentiate into chondrocytes in vitro in the presence or absence of transforming growth factor beta (TGF-ß1). Human BMSCs and AMSCs were collected from healthy donors during orthopaedic surgeries, in vitro cultured in three passages to obtain the required quantity of cells. A pellet culture system was used for chondrocyte differentiation.
At 21 days of cultivation, cell aggregates grown in the chondrogenic medium were larger than those cultured in the control medium. Both the BMSCs and AMSCs pellet cultures showed spontaneous chondrogenesis. Histological staining with haematoxylin and eosin and Masson's trichrome stains, as well as immunohistochemical staining to detect type II collagen revealed no apparent differences between the pellet cultures with TGF-ß1 presence and those without it. The real-time RT-PCR detected expression of the type II collagen gene in all tested cultures. In the BMSCs pellet culture only, TGF-ß1 presence resulted in a decrease in type I collagen mRNA levels and in an increase in type II collagen mRNA values.
Our results showed an in vitro chondrogenic potential of mature human mesenchymal stem cells derived from both bone marrow and adipose tissue. In agreement with the relevant literature data, we suggest that both cell types have an equal prospect for use in cartilage tissue engineering.
许多先天性和后天性疾病以及损伤后遗症都与关节软骨退变有关,这对患者的生活质量产生不利影响。目前使用培养软骨细胞的细胞疗法存在缺点,因为在培养过程中软骨细胞会发生去分化过程。我们认为间充质干细胞疗法提供了新的治疗选择。
用于软骨细胞分化的成人间充质干细胞(MSCs)通常从骨髓间充质干细胞(BMSCs)中获取。在本研究中,将这些细胞与源自脂肪组织的间充质干细胞(AMSCs)进行比较。研究目的是验证人BMSCs和AMSCs在有或没有转化生长因子β(TGF-β1)存在的情况下在体外分化为软骨细胞的能力。人BMSCs和AMSCs在骨科手术期间从健康供体中采集,在体外培养三代以获得所需数量的细胞。采用微团培养系统进行软骨细胞分化。
培养21天时,在软骨形成培养基中生长的细胞聚集体比在对照培养基中培养的细胞聚集体大。BMSCs和AMSCs的微团培养均显示出自发性软骨形成。苏木精和伊红染色以及Masson三色染色的组织学染色,以及检测II型胶原蛋白的免疫组织化学染色显示,在有TGF-β1存在和不存在TGF-β1的微团培养之间没有明显差异。实时RT-PCR检测到所有测试培养物中II型胶原蛋白基因的表达。仅在BMSCs微团培养中,TGF-β1的存在导致I型胶原蛋白mRNA水平降低,II型胶原蛋白mRNA值增加。
我们的结果显示了源自骨髓和脂肪组织的成熟人间充质干细胞的体外软骨形成潜力。与相关文献数据一致,我们认为这两种细胞类型在软骨组织工程中的应用前景相同。